The labeling of 3T3-L1 preadipocyte cells with enhanced green fluorescent protein.
- Author:
Cheng-Jian LI
1
;
Jun-Ying CHENG
;
Xiao-Lan ZHANG
;
Chong-Ben ZHANG
Author Information
1. College of life Sciences, Peking University, Beijing 100871, China.
- Publication Type:Journal Article
- MeSH:
3T3-L1 Cells;
Adipocytes;
cytology;
Animals;
Cell Differentiation;
Green Fluorescent Proteins;
genetics;
Mice;
PPAR gamma;
genetics;
Promoter Regions, Genetic;
Stem Cells;
cytology
- From:
Chinese Journal of Biotechnology
2004;20(4):607-609
- CountryChina
- Language:English
-
Abstract:
A cell model is desired for adipocyte differentiation investigation and for high-throughput screening of anti-obesity and anti-diabetes molecules from chemical resources due to the world wide epidemic of obesity and diabetes. In order to establish such a cell model, a plasmid of pPPARgamma2-promoter-EGFP was constructed by inserting a 660bp sequence of mouse PPARgamma2 promoter into the Ase I and Kpn I sites of pEGFP-N3 and transferred into 3T3-L1 preadipocyte cells. The cells were induced to differentiate and the expression of PPARgamma2 was detected by the microscopic observation of EGFP and by RT-PCR assays. The results showed that the EGFP gene expression patterns were similar to that of pPPARgamma2's, which indicated that the EGFP gene was transferred into the mouse 3T3-L1 preadipocyte cells, and its expression was under the control of pPPARgamma2 promoter. RT-PCR assays showed that the EGFP expression authentically represented the stable expression of PPARgamma2. In conclusion, a preadipocyte cell line expressing EGFP under the control of the promoter of adipocyte-specific expression gene PPARgamma2 was generated. The cell line provides a powerful approach for the research of adipocyte differentiation and for the high-throughput screening of anti-obesity and anti-diabetes chemicals.
